The ALOX12 Knockout KYSE-150 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell pool constructed from the human KYSE-150 esophageal squamous cell carcinoma (ESCC) line. This mixed-genotype population features targeted disruption of the ALOX12 gene, yielding a loss-of-function model that captures allelic diversity without single-cell cloning. The polyclonal format minimizes clonal artifacts and preserves the heterogeneity inherent to cancer cell populations, making it a robust tool for investigating arachidonate 12-lipoxygenase function in tumor biology.
The parental KYSE-150 cell line was established from a poorly differentiated esophageal squamous cell carcinoma of a 49-year-old Japanese male. These adherent epithelial cells retain characteristics of the original tumor and are extensively employed in ESCC studies, including investigations of invasion, drug resistance, and signal transduction. The cell line??s genetic background, which harbors TP53 mutations and other oncogenic alterations, provides a clinically relevant context for functional genomics. By introducing ALOX12 knockout into this well-characterized model, the product enables precise dissection of lipid mediator signaling in esophageal cancer.
ALOX12 encodes arachidonate 12-lipoxygenase, a rate-limiting enzyme that converts arachidonic acid to 12-HPETE, which is rapidly reduced to the bioactive lipid mediator 12-HETE. This eicosanoid acts through the G-protein-coupled receptor GPR31 to stimulate downstream transcription factors NF-??B and AP-1, promoting pro-inflammatory and pro-survival gene expression. ALOX12 activity is regulated by upstream MAPK/ERK and protein kinase C (PKC) cascades, and it functionally interacts with GPX4 to influence ferroptosis sensitivity by modulating lipid peroxidation. In ESCC cells, ALOX12-derived 12-HETE drives production of cytokines such as IL-6 and TNF??, contributing to a tumor-supportive microenvironment and resistance to oxidative stress.
Disruption of ALOX12 in KYSE-150 cells abolishes 12-HETE synthesis, thereby blocking GPR31 receptor activation and attenuating downstream pro-inflammatory signaling cascades. This loss-of-function model is particularly germane to ESCC research, where elevated ALOX12 expression has been correlated with poor patient prognosis and enhanced tumor cell proliferation. The polyclonal knockout pool permits rigorous evaluation of how ALOX12 ablation impacts hallmark cancer cell behaviors, including anchorage-independent growth, migratory capacity, and susceptibility to ferroptotic death. Given the established interplay between lipid peroxidation and ferroptosis execution, this model is well-suited for investigating whether pharmacological or genetic inhibition of ALOX12 can sensitize esophageal carcinoma cells to oxidative overload and cell death.
This polyclonal knockout product supports target validation, eicosanoid signaling studies, and ferroptosis research. Compatible assays include Western blot for ALOX12, LC-MS/MS for 12-HETE, MTS and Transwell assays, and lipid peroxidation readouts via BODIPY-C11 or GPX4 immunoblotting. RT-qPCR measures NF-??B targets IL-6 and TNF??, and flow cytometry assesses apoptosis. The pooled cells provide a reliable source for inhibitor screens. For protocols or support, contact Ascent Research.
